Morphological Analysis of Dopaminergic Neurons with Age Using Caenorhabditis elegans GFP Reporter Strains

Materials and Reagents

1. Plastic plates , 60 x 15 mm (Olympus Plastics, catalog number: 32-105G )
   
2. Microscopic slides (Denville Scientific, catalog number: M1002 )
   
3. Micro cover glass, 22 x 22 mm (VWR, catalog number: 48366-067 )
   
4. OP50-1 Bacteria (E. coli https://cgc.umn.edu/strain/OP50-1)
   
5. C. elegans reporter strains
   BY200 - Pdat-1::GFP(vtIs1) V (Michael Aschner, Albert Einstein College of Medicine, NY, USA)
   BY250 - Pdat-1::GFP(vtIs7) V (Randy Blakely, Florida Atlantic University, FL, USA)
   UA44 - [baInl1; Pdat-1::α-syn high, Pdat-1::gfp] (Guy Caldwell, University of Alabama, AL, USA)
   UA57 - baIs4 [dat-1p::GFP + dat-1p::CAT-2] (CGC, University of Minnesota, MN, USA)
   
6. Sodium hypochlorite solution (Avantor Performance Materials, J.T. Baker, catalog number: 9416-01 )
   
7. M9 buffer
8. NGM agar
   
9. Agarose LE (Apex Chemicals and Reagents, catalog number: 01132-34 )
   
10. Levamisole hydrochloride (MedChemExpress, catalog number: HY-13666 )
    
11. Immersion Oil (Immersol 518F, Carl Zeiss, catalog number: 444960-0000-000 )
    

Equipment

1. Centrifuge
   
2. Shaker
   
3. Fluorescent Microscope (Olympus, model: BX51 , TRF)
   
4. Digital camera (Hamamatsu ORCA-ER, version: C4742-80)
   

Software

1. Image J
   
2. GraphPad Prism
   

Procedure